LOCUS       sequence-384000-        4459 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro transcription of mKate2 tagged zebrafish caveolin1 from the
            T3 promoter. The red fluorescent tag is at the N-terminus. Parton 
            lab clone KZW.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4459)
  AUTHORS   Wu Y, Lim YW, Stroud DA, Martel N, Hall TE, Lo HP, Ferguson C, Ryan 
            MT, McMahon KA, Parton RG
  TITLE     Caveolae sense oxidative stress through membrane lipid peroxidation 
            and cytosolic release of CAVIN1 to regulate NRF2.
  JOURNAL   Dev Cell. 2023 Mar 13;58(5):376-397.e4. doi: 
            10.1016/j.devcel.2023.02.004. Epub 2023 Feb 28.
  PUBMED    36858041
REFERENCE   2  (bases 1 to 4459)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.devcel.2023.02.004"; journalName: "Dev Cell"; date: 
            "2023-03-13- 13"; volume: "58"; issue: "5"; pages: "376-397.e4"
COMMENT     Sequence Label: mKate2-DrCav1 in pT3TS-Dest
FEATURES             Location/Qualifiers
     source          1..4459
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     5'UTR           44..86
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     protein_bind    93..117
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      118..127
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             124..819
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     protein_bind    complement(821..845)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1390..1410)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     3'UTR           1435..1560
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(1647..1666)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1648..1666)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1673..1690)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1673..1689)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1682..1704)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(1831..2286)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1968..1989)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2180..2199
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2313..2417
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2418..2486,2487..3278)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2636..2655)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3449..4037
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3938..3957
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4191..4208
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4325..4346
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4361..4366,4367..4384,4385..4391)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4399..4415
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     4404..4426
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4423..4439
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4423..4439
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aattaaccct cactaaaggg aacaaaagct ggagctaagc ttgcttgttc tttttgcaga
       61 agctcagaat aaacgctcaa ctttggcaga tctcaagttt gtacaaaaaa gcaggctgcc
      121 accatggtga gcgagctgat taaggagaac atgcacatga agctgtacat ggagggcacc
      181 gtgaacaacc accacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc
      241 cagaccatga gaatcaaggc ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg
      301 gctaccagct tcatgtacgg cagcaaaacc ttcatcaacc acacccaggg catccccgac
      361 ttctttaagc agtccttccc cgagggcttc acatgggaga gagtcaccac atacgaagac
      421 gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct catctacaac
      481 gtcaagatca gaggggtgaa cttcccatcc aacggccctg tgatgcagaa gaaaacactc
      541 ggctgggagg cctccaccga gaccctgtac cccgctgacg gcggcctgga aggcagagcc
      601 gacatggccc tgaagctcgt gggcgggggc cacctgatct gcaacttgaa gaccacatac
      661 agatccaaga aacccgctaa gaacctcaag atgcccggcg tctactatgt ggacagaaga
      721 ctggaaagaa tcaaggaggc cgacaaagag acctacgtcg agcagcacga ggtggctgtg
      781 gccagatact gcgacctccc tagcaaactg gggcacagag acccagcttt cttgtacaaa
      841 gtggtgacta gcggatacaa ggacgggaca cctgaagagg aatacgctca ctcgccgttt
      901 atcaggaaac aggggaacat ttacaaacca aacaataaag aaatggataa cgacagcatc
      961 aacgaaaaga cacttcagga tgtccacacc aaggagattg acctggtcaa ccgagaccca
     1021 aagcatttaa atgacgatgt ggtgaaggtg gactttgagg acgtgatcgc cgagcctgcc
     1081 ggcacctaca gcttcgacgg cgtgtggaag gcgagcttca ccaccttcac agtaaccaaa
     1141 tattggtgct acaggctgct gacagcgctg gtgggcatcc cactcgccct ggtatggggc
     1201 atcttcttcg ccatcctctc cttcatccac atctgggccg tggtgccttg cgtgaagagc
     1261 tacctaatcg agatccactg catcagtcga gtttactcca tctgtgtgca caccttctgc
     1321 gacccactct ttgaggccat ggggaaatgc tttagtaacg tccgggtcac tgctactaag
     1381 gtggtgtaac aactttatta tacatagttg ctcgagcgga tctggttacc actaaaccag
     1441 cctcaagaac acccgaatgg agtctctaag ctacataata ccaacttaca ctttacaaaa
     1501 tgttgtcccc caaaatgtag ccattcgtat ctgctcctaa taaaaagaaa gtttcttcac
     1561 attctaaaaa aaaaaaaaaa aaaaaaaaaa aaaacccccc ccccccccct gcagggccaa
     1621 gtcggcctct agaggtaccc aattcgccct atagtgagtc gtattacaat tcactggccg
     1681 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     1741 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     1801 aacagttgcg cagcctgaat ggcgaatggc aaattgtaag cgttaatatt ttgttaaaat
     1861 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     1921 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     1981 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     2041 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     2101 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     2161 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     2221 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     2281 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2341 aaatacattc aaatatgtat ctgctcatga gacaataacc ctgataaatg cttcaataat
     2401 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2461 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2521 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2581 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     2641 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     2701 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     2761 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     2821 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2881 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2941 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     3001 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     3061 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3121 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3181 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3241 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3301 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3361 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3421 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3481 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3541 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     3601 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3661 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3721 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3781 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3841 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3901 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3961 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4021 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4081 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4141 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     4201 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     4261 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     4321 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     4381 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     4441 atgattacgc caagctcga
//
